Wide and high resolution tension measurement using FRET in embryo.
Wide and high resolution tension measurement using FRET in embryo.
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DOI：
10.1038/srep28535
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发表时间：
2016-06-23
影响因子：
4.6
通讯作者：
Michiue T
中科院分区：
文献类型：
--
作者：
Yamashita S;Tsuboi T;Ishinabe N;Kitaguchi T;Michiue T
在胚胎发育过程中，物理力在形态发生和分化中起着重要作用。拉伸敏感荧光共振能量转移（FRET）有可能提供非侵入性的张力测量活组织内。在这项研究中，我们引入了一个基于FRET的辅肌动蛋白张力传感器非洲爪蟾胚胎，并证明了这种传感器捕获跨分化外胚层的张力变化。肌动蛋白张力传感器，含有mCherry和EGFP连接的蜘蛛丝蛋白，在人胚肾（HEK）细胞和胚胎中进行了验证。它与肌动蛋白丝共定位，并改变FRET效率响应肌动蛋白丝破坏，肌球蛋白失活，和渗透扰动。延时FRET分析表明，在原肠胚和神经胚形成过程中，神经外胚层比表皮外胚层承受更高的张力，细胞的形态发生行为与张力差异相关。这些数据证实，传感器使我们能够同时测量跨组织的张力，并具有高分辨率。
During embryonic development, physical force plays an important role in morphogenesis and differentiation. Stretch sensitive fluorescence resonance energy transfer (FRET) has the potential to provide non-invasive tension measurements inside living tissue. In this study, we introduced a FRET-based actinin tension sensor into Xenopus laevis embryos and demonstrated that this sensor captures variation of tension across differentiating ectoderm. The actinin tension sensor, containing mCherry and EGFP connected by spider silk protein, was validated in human embryonic kidney (HEK) cells and embryos. It co-localized with actin filaments and changed FRET efficiencies in response to actin filament destruction, myosin deactivation, and osmotic perturbation. Time-lapse FRET analysis showed that the prospective neural ectoderm bears higher tension than the epidermal ectoderm during gastrulation and neurulation, and cells morphogenetic behavior correlated with the tension difference. These data confirmed that the sensor enables us to measure tension across tissues concurrently and with high resolution.